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HUMAN EMBRYONIC STEM CELLS
AND ITS POSSIBLE
APPLICATIONS IN GENETICS
Professor ANDRE VAN STEIRTEGHEM
Koninklijke Academie voor Geneeskunde van België
Emeritus Professor Vrije Universiteit Brussel
Honorary Consultant Centre for Reproductive Medicine, Universitair
Ziekenhuis Brussel
Editor-in-Chief Human Reproduction
22-5-2017
Herhaling titel van presentatie
FEAM 2007
1
Outline lecture
•
Introduction
•
Legal and ethical framework
•
Derivation and differentiation of hESC lines
•
hESC lines from genetically abnormal
embryos
•
European registry of hESC lines
•
Conclusions
•
Acknowledgments
Introduction
•
Definition of human Embryonic Stem Cells
(hESC)
• Derived from preimplantation embryo
• Proliferate without differentiation for a long
period
• Have potential of stable differentiation
towards three embryonic layers and all
types of cells of an embryo and an adult
individual
Derivation of a hES line
Opinion poll after first publication
ESC for therapy
IVF for infertility
USA
68%
81%
UK
62%
62%
Australia
72%
86%
Great “hope” for stem cell therapy
Outline lecture
•
Introduction
•
Legal and ethical framework
•
Derivation and differentiation of hESC lines
•
hESC lines from genetically abnormal
embryos
•
European registry of hESC lines
•
Conclusions
•
Acknowledgments
Legal and ethical issues
•
Great variety of legal framework regarding
hESC
•
Liberal law: UK, Belgium
•
Restrictive law: Germany, Italy
•
Intermediate law: France, The Netherlands
•
Particular situation in the USA
•
Approval of ethical committee is always
required (local or national)
hES CELLS IN BELGIUM
•
Belgian law on the protection of human
embryo in-vitro allows research on IVF-ICSI-
PGD research embryos
•
If necessary embryos can be made for
research
•
Stem cell research and therapeutic cloning is
allowed
Regulations in European Member States Regarding Human Embryonic
Stem Cell Research (July 2003)
Policy
EU Member States
No specific legislation regarding human embryo research
Luxembourg, Portugal
Allowing for the creation of human embryos for stem cell
procurement by law
Belgium, United Kingdom
Prohibition of the procurement of embryonic stem cells from
human embryos
Austria, Spain, France, Ireland, Italy
Prohibition of the procurement of embryonic stem cells from
human embryos but allowing by law for the importation of human
embryonic stem cell lines
Germany
Allowing for the procurement of human embryonic stem cells from
supernumerary embryos by law
Belgium, Denmark, Finland, Greece,
Netherlands, Sweden, United Kingdom
Prohibition of the creation of human embryos for research purposes
and for the procurement of stem cells by law or by ratification of
the Convention of the Council of Europe on Human Rights and
Biomedicine signed in Oviedo on April 4, 1997
Austria, Denmark, Germany, Spain,
Finland, France, Greece, Ireland,
Netherlands, Portugal
Outline lecture
•
•
•
•
•
•
•
Introduction
Legal and ethical framework
Derivation and differentiation of hESC
lines
hESC lines from genetically abnormal
embryos
European registry of hESC lines
Conclusions
Acknowledgments
Where do human embryonic stem cells
come from?
•
ICM of expanded blastocysts
•
Put in culture on mouse embryonic
fibroblasts
•
Culture medium contains fetal calf
serum/serum replacement and BFGF
Derivation of hESC
5-day old plated ICM
Culture of hESC
ES cell colonies, > 30 passages
Propagation of hESC cells
Propagation by cutting
Totipotency of hESC: teratoma in vivo
Teratoma VUB01
Primitive bronchial epithelium
Chondroid tissue
Primitive nerve fibers
Totipotency of hESC: embryoid bodies in
vitro
• Clumps of hESC in
suspension
• No MEF, no BFGF
• Spontaneously differentiate
to all three germ layers
• Classic method used for
differentiation in the mouse
• Plate the cells
• Look for the interesting
cells
Directed differentiation: in vivo situation
Embryonic
ectoderm
Embryonic
epiblast
Epiblast
ICM
Blastocyst
Hypoblast
Trophoblast
Amnionic
ectoderm
Embryonic
endoderm
Primitive
streak
Embryonic
mesoderm
Outline lecture
•
Introduction
•
Legal and ethical framework
•
Derivation and differentiation of hESC lines
•
hESC lines from genetically abnormal
embryos
•
European registry of hESC lines
•
Conclusions
•
Acknowledgments
•
PGD introduced in 1990 can be carried out
for an increasing number of monogenic and
chromosomal disorders for couples with a
high risk of an affected child
•
Affected embryos will not be transferred
and may be used to derive human
embryonic stem cell lines if there are no
legal restrictions
•
Belgium law (May 2003) on the protection
of human embryo in vitro allows embryo
research and derivation of hESC lines
•
Different hES cell lines have different gene
expression characteristics  comparative
studies are required before considering
clinical use
•
Derivation of most cell lines using MEF 
only for in vitro research
•
Feeder-free conditions reported
(Klimanskaya et al. Lancet 365: 1636,
2005)
•
Sofar at Vrije Universiteit Brussel 14 well
established hESC lines
•
Four from genetically normal embryos
•
Ten from genetically abnormal PGD embryos:
myotonic dystrophy, carrier cystic fibrosis,
Huntington disease, Marfan syndrome, fascioscapulo-humeral disease, spinocerebellar ataxia,
Fragile-X syndrome (2 lines), adult polykystic
kidney disease and bare lymphocyte syndrome
•
•
•
•
•
Genetically normal and affected hES cell
lines can be compared
DM1, CF and HD common genetic diseases
without treatment
Model for human diseases: hES cell lines
can be unlimited source of specific cell
types involved in pathology of the disease
eg CNS neurons in HD
Study of mechanisms of dynamic mutations
caused by unstable triplet repeats using
hES cell lines from DM1 and HD
CF line with F508 and 5T pathogenesis of
CBAVD
•
•
•
•
Each of the genetic diseases are very rare
(< 5 births per 10,000)  maximum of 200
000 affected patients per disease in
European Union
However, there are about 7000 different
genetic diseases  the number of affected
patients is high (estimate in Europe of 25-30
million)
Pharmacotherapy for genetic diseases is
almost non-existing; validation of new
compounds is currently almost impossible
Search for biomarkers associated with the
expression of mutated genes
•
Functional genomic studies can be
carried out to search for the molecular
mechanisms of the genetic diseases
•
Using chemical data bases it may be
possible to discover molecules with
therapeutic potential
Outline lecture
•
Introduction
•
Legal and ethical framework
•
Derivation and differentiation of hESC lines
•
hESC lines from genetically abnormal
embryos
•
European registry of hESC lines
•
Conclusions
•
Acknowledgments
•
•
•
•
•
•
European Commission announced creation of a
European registry for human embryonic stem cell
research
To promote coordination of stem cell research in
Europe
Detailed information on all embryonic stem cell
lines available in Europe
Registry available to public via internet:
characteristics, contact data, interesting
developments (clinical trials and hESC lines from
Eu-funded projects
Advisory Board, Steering Committee and
independent Ethics Advisory Board
Stem cell research funded by Sixth and Seventh
Framework Programme
Outline lecture
•
Introduction
•
Legal and ethical framework
•
Derivation and differentiation of hESC lines
•
hESC lines from genetically abnormal
embryos
•
European registry of hESC lines
•
Conclusions
•
Acknowledgments
Outline lecture
•
Introduction
•
Legal and ethical framework
•
Derivation and differentiation of hESC lines
•
hESC lines from genetically abnormal
embryos
•
European registry of hESC lines
•
Conclusions
•
Acknowledgments
Collaborators
Prof. Dr. I. Liebaers
Prof. Dr. K. Sermon
Prof. Dr. J. Vanderelst
Prof. Dr. P. Devroey
Dr. M. De Rycke
Dr. H. Van de Velde
Stem cell team:
Claudia Spits
Ileana Mateizel
Nele De Temmerman
Urielle Ullmann
Mieke Geens
Rogier Schellens
Lindsay Van Haute